Quantitative, chemically specific images of biological systems would be invaluable in unraveling the bioinorganic chemistry of biological tissues. Here we report the spatial distribution and chemical forms of selenium in Astragalus bisulcatus (two-grooved poison or milk vetch), a plant capable of accumulating up to 0.65% of its shoot dry biomass as Se in its natural habitat. By selectively tuning incident x-ray energies close to the Se K-absorption edge, we have collected quantitative, 100-m-resolution images of the spatial distribution, concentration, and chemical form of Se in intact root and shoot tissues. To our knowledge, this is the first report of quantitative concentration-imaging of specific chemical forms. Plants exposed to 5 M selenate for 28 days contained predominantly selenate in the mature leaf tissue at a concentration of 0.3-0.6 mM, whereas the young leaves and the roots contained organoselenium almost exclusively, indicating that the ability to biotransform selenate is either inducible or developmentally specific. While the concentration of organoselenium in the majority of the root tissue was much lower than that of the youngest leaves (0.2-0.3 compared with 3-4 mM), isolated areas on the extremities of the roots contained concentrations of organoselenium an order of magnitude greater than the rest of the root. These imaging results were corroborated by spatially resolved x-ray absorption near-edge spectra collected from selected 100 ؋ 100 m 2 regions of the same tissues.
Selenium is now known to play essential metabolic roles in most or perhaps all eukaryotes (1), but its toxic properties were recognized earlier in the poisoning of range animals eating selenium-hyperaccumulating plants (2). Hyperaccumulating plants actively and specifically accumulate high levels of metal or metalloid compounds into their tissues (3), and there is considerable interest in the biochemical mechanisms of hyperaccumulation, particularly with regard to potential phytoremediation strategies for contaminated environments (4) and for use as dietary supplements or ''nutraceuticals'' (5). The two-grooved milk-or poison-vetch, Astragalus bisulcatus, also known by cattle ranchers as ''locoweed,'' hyperaccumulates up to 0.65% of its shoot dry biomass as Se (6) from inorganic selenium naturally present in soil in parts of its range. The major Se-containing compound in the leaves is Se-methylselenocysteine, with smaller amounts of selenocystathione and ␥-glutamyl-Se-methylselenocysteine localized in the seed pods (7). Other organoselenium compounds are also produced but in smaller amounts (dimethyldiselenide in particular is clearly indicated by its characteristic odor). The purification, cloning, and characterization of a specific selenocysteine methyltransferase involved in synthesizing Se-methylselenocysteine from selenocysteine and S-methylmethionine has recently been reported (8). Despite this progress, much remains to be learned about the details of Se uptake, transport, and storage in hyperaccumulating plants. X-ray ...